Salts of 2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1h)-one derivatives having both muscarinic receptor antagonist and beta2 adrenergic receptor agonist activities

ABSTRACT

The present invention is directed to crystalline addition salts of (ii) 8-hydroxyquinolin-2(1H)-one derivatives and (ii) a dicarboxylic acid or a sulfimide, or a pharmaceutically acceptable solvates thereof.

FIELD OF THE INVENTION

The present invention is directed to a pharmaceutically acceptablecrystalline addition salt of (i) a2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1H)-one derivative and (ii) adicarboxylic acid or a sulfimide derivative, or a pharmaceuticallyacceptable solvate thereof. The invention is also directed topharmaceutical compositions comprising the salts, methods of using themto treat respiratory diseases associated with dual muscarinic receptorantagonist and β2 adrenergic receptor agonist activities, and processesand intermediates useful for preparing such salts.

BACKGROUND OF THE INVENTION

WO 2013/068552 and WO 2013/068554 disclose compounds which are known tohave a dual muscarinic receptor antagonist and β2 adrenergic receptoragonist activity. However, many of these compounds cannot be formulatedfor effective delivery by inhalation as a dry powder which ischallenging. It requires careful control of the particle size of thepowder which is to be inhaled, and careful control of the particle sizedistribution. Further, it is important to avoid particle agglomerationor aggregation. In addition, when preparing pharmaceutical compositionsand formulations for use in such devices, it is highly desirable to havea crystalline form of a therapeutic agent that is neither hygroscopicnor deliquescent and which has a relatively high melting point therebyallowing the material to be micronized without significant decompositionor loss of crystallinity.

Although the 2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1H)-onederivatives disclosed in WO 2013/068552 and WO 2013/068554 have shownadequate pharmacological behavior, it has proved difficult to obtainthem in the form of a salt which is crystalline, not hygroscopic nordeliquescent and which has a relatively high melting point to enablemicronization.

So far no crystalline salt of any of the compounds disclosed in WO2013/068552 and WO 2013/068554 having the desired properties has beenreported.

Accordingly, a need exists for stable, non-deliquescent salt forms of atleast some of these compounds having acceptable levels of hygroscopicityand relatively high melting points.

SUMMARY OF THE INVENTION

The present invention provides a pharmaceutically acceptable crystallineaddition salt of (i) a2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1H)-one derivative and (ii) adicarboxylic acid or a sulfimide derivative, or a pharmaceuticallyacceptable solvate thereof, wherein the2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1H)-one derivative is acompound of formula(I):

Wherein:

-   -   R¹, R² and R³ independently represent a hydrogen atom or a C₁₋₂        alkyl group,    -   R⁴ represents a hydrogen atom, a hydroxy group, a hydroxymethyl        group or a linear or branched C₁₋₄ alkyl group,    -   R⁵ and R⁶ independently represents a thienyl group, a phenyl        group, a benzyl group or a C₄₋₆ cycloalkyl group,    -   V and W are independently selected from a —N—, —S— and —C(O)—        group,    -   n and m independently have a value from 0 to 4.

The dash line (----) in formula(I) means that the bond may be either asingle bond or a double bond.

The invention also provides a pharmaceutical composition comprising atherapeutically effective amount of a salt of the invention and apharmaceutically acceptable carrier.

The invention further provides a combination comprising a salt of theinvention and one or more other therapeutic agents.

The invention also provides a salt of the invention, a pharmaceuticalcomposition of the invention or a combination of the invention, for usein the treatment of a pathological condition or disease associated withboth β2 adrenergic receptor agonist and muscarinic receptor antagonistactivity.

The invention further provides the use of a salt of the invention, apharmaceutical composition of the invention or a combination of theinvention, in the manufacture of a medicament for the treatment of apathological condition or disease associated with both β2 adrenergicreceptor agonist and muscarinic receptor antagonist activity.

The invention also provides a method for treating a subject afflictedwith a pathological condition or disease associated with both β2adrenergic receptor agonist and M3 muscarinic receptor antagonistactivity, which comprises administering to said subject an effectiveamount of a salt of the invention, a pharmaceutical composition of theinvention or a combination of the invention.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 shows the Powder X-Ray Diffraction (PXRD) pattern fortrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]cyclohexyl hydroxy(di-2-thienyl)acetate saccharinate.

FIG. 2 shows the ¹H-NMR (500 MHz MeOD-d4) fortrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]cyclohexyl hydroxy(di-2-thienyl)acetate saccharinate.

FIG. 3 shows the Thermogravimetric (TG) analysis and the differentialscanning calorimetry (DSC) analysis oftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]cyclohexyl hydroxy(di-2-thienyl)acetate saccharinate.

FIG. 4 shows the powder X-ray diffraction (PXRD) pattern fortrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate.

FIG. 5 is the ¹H-NMR spectrum (400 MHz, d6-DMSO) fortrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate.

FIG. 6 shows the DSC analysis fortrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate

FIG. 7 shows the TG analysis fortrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate.

DETAILED DESCRIPTION OF THE INVENTION

When describing the salts, compositions and methods of the invention,the following terms have the following meanings, unless otherwiseindicated.

The term “therapeutically effective amount” refers to an amountsufficient to effect treatment when administered to a patient in need oftreatment.

The term “treatment” as used herein refers to the treatment of a diseaseor medical condition in a human patient which includes:

-   -   (a) preventing the disease or medical condition from occurring,        i.e., prophylactic treatment of a patient;    -   (b) ameliorating the disease or medical condition, i.e., causing        regression of the disease or medical condition in a patient;    -   (c) suppressing the disease or medical condition, i.e., slowing        the development of the disease or medical condition in a        patient; or    -   (d) alleviating the symptoms of the disease or medical condition        in a patient.

The phrase “disease or condition associated with β2 adrenergic receptoragonist and M3 muscarinic receptor antagonist activities” includes alldisease states and/or conditions that are acknowledged now, or that arefound in the future, to be associated with both β2 adrenergic receptoragonist and M3 muscarinic receptor antagonist activity. Such diseasestates include, but are not limited to, pulmonary diseases, such asasthma and chronic obstructive pulmonary disease (including chronicbronchitis and emphysema), as well as neurological disorders and cardiacdisorders. β2 adrenergic receptor activity is also known to beassociated with pre-term labor (see International Patent ApplicationPublication Number WO 98/09632), glaucoma and some types of inflammation(see International Patent Application Publication Number WO 99/30703 andPatent Application Publication Number EP 1 078 629).

On the other hand M3 receptor activity is associated withgastrointestinal-tract disorders such as Irritable bowel syndrome (IBS)(see, for ex., U.S. Pat. No. 5,397,800), gastrointestinal (GI) ulcers,spastic colitis (see, for ex., U.S. Pat. No. 4,556,653); urinary-tractdisorders such as urinary incontinence (see, for ex., J.Med.Chem., 2005,48, 6597-6606), pollakiuria; motion sickness and vagally induced sinusbradycardia.

The compounds of the invention may exist in both unsolvated and solvatedforms. The term solvate is used herein to describe a molecular complexcomprising a compound of the invention and an amount of one or morepharmaceutically acceptable solvent molecules. The term hydrate isemployed when said solvent is water. Examples of solvate forms include,but are not limited to, compounds of the invention in association withwater, acetone, dichloromethane, 2-propanol, ethanol, methanol,dimethylsulfoxide (DMSO), ethyl acetate, acetic acid, ethanolamine, ormixtures thereof. It is specifically contemplated that in the presentinvention one solvent molecule can be associated with one molecule ofthe compounds of the present invention, such as a hydrate.

Furthermore, it is specifically contemplated that in the presentinvention, more than one solvent molecule may be associated with onemolecule of the compounds of the present invention, such as a dihydrate.Additionally, it is specifically contemplated that in the presentinvention less than one solvent molecule may be associated with onemolecule of the compounds of the present invention, such as ahemihydrate. Furthermore, solvates of the present invention arecontemplated as solvates of compounds of the present invention thatretain the biological effectiveness of the non-solvate form of thecompounds.

As used herein the term C₁₋₄ alkyl embraces linear or branched radicalshaving 1 to 4 carbon atoms. Examples include methyl, ethyl, propyl,butyl, isopropyl and tert-butyl radicals.

As used herein the term C₁₋₂ alkyl embraces radicals having 1 to 2carbon atoms. Examples include methyl and ethyl radicals.

As used herein, the term C₄-C₆ cycloalkyl radical embraces saturatedmonocyclic carbocyclic radicals having from 4 to 6 carbon atoms.Examples of monocyclic cycloalkyl groups include cyclobutyl, cyclopentyland cyclohexyl group.

Typically, in the compound of formula (I), R¹, R² and R³ independentlyrepresent a hydrogen atom or a methyl group, preferably both R¹ and R²are a hydrogen atom while R₃ represents a methyl group.

Typically, in the compound of formula (I), V represents a —N— or —S—group, and W represents —N— or —C(O)— group.

Typically, in the compound of formula (I), n has a value of 0 or 1, andm has a value of 2 or 3. Preferably, n has a value of 0 while m has avalue of 3.

Preferably, in the compound of formula (I), both R¹ and R² are ahydrogen atom, R₃ represents a methyl group, V represents a —N— or —S—group and W represents —N— or —C(O)— group, n has a value of 0 and m hasa value of 3.

Typically, the dicarboxylic acid is selected from the group consistingof L-tartaric acid, fumaric acid, succinic acid, maleic acid and oxalicacid, preferably fumaric acid

Typically, the sulfimide derivative is selected from the groupconsisting of benzoic sulfimide (also known as saccharin),thieno[2,3-d]isothiazol-3(2H)-one 1,1-dioxide and isothiazol-3(2H)-one1,1-dioxide, preferably benzoic sulfimide (saccharin).

In a preferred embodiment, the dicarboxylic acid is fumaric acid and thesulfimide derivative is saccharin.

Particular individual salt compounds of the invention include:

-   -   trans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]        cyclohexyl hydroxy(di-2-thienyl)acetate saccharinate, and    -   trans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)-amino]cyclohexyl        hydroxy(di-2-thienyl)acetate fumarate,

General Synthetic Procedures

The salts of the invention can be prepared using the methods andprocedures described herein, or using similar methods and procedures. Itwill be appreciated that where typical or preferred process conditions(i.e., reaction temperatures, times, mole ratios of reactants, solvents,pressures, etc.) are given, other process conditions can also be usedunless otherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Processes for preparing salts of the invention are provided as furtherembodiments of the invention and are illustrated by the proceduresbelow.

The salts of the invention can be synthesized from compounds of formula(I) and from the appropriate dicarboxylic acid or sulfimide derivatives,which will generally be commercially available from, for example,Aldrich.

Suitable solvents for carrying out the reaction can be selected by askilled chemist and may depend on the specific salt to be formed.Mixtures of appropriate solvents may be used, optionally containingwater. For example, the appropriate solvents may be selected fromethanol, acetone, ethyl acetate, tetrahydrofuran or a mixture thereof.

Upon completion of any of the foregoing reactions, the salt can beisolated from the reaction mixture by any conventional means such asprecipitation, concentration, centrifugation and the like.

It will be appreciated that while specific process conditions (i.e.reaction temperatures, times, mole ratios of reactants, solvents,pressures, etc.) are given, other process conditions can also be usedunless otherwise stated.

To prepare the salts of the present invention, the free base istypically dissolved in an appropriate solvent which in some examples isheated to approximately 60-80° C. Then a solution of the appropriatedicarboxylic acid or a sulfimide in an suitable solvent, preferably thesame solvent as that in which the free base is dissolved, is typicallyadded to the heated solution. The mixture is then optionally stirred for15-300 minutes at 60-80° C. or at room temperature. The mixture is thentypically cooled, for example down to 20-25° C. The precipitate formedis isolated by filtration, washed with an appropriate solvent and driedfor example in vacuum.

Pharmaceutical Compositions

The invention also encompasses pharmaceutical compositions comprising atherapeutically effective amount of a salt of the invention or anenantiomer or pharmaceutically acceptable solvate thereof and apharmaceutically acceptable carrier. Typically the pharmaceuticalcomposition is formulated for administration by inhalation, preferablyas a dry powder.

Typically, the pharmaceutical composition further comprises atherapeutically effective amount of one or more other therapeuticagents.

The pharmaceutical formulations may conveniently be presented in unitdosage form and may be prepared by any of the methods well known in theart of pharmacy. All methods include the step of bringing the activeingredient(s) into association with the carrier. In general theformulations are prepared by uniformly and intimately bringing intoassociation the active ingredient with liquid carriers or finely dividedsolid carriers or both and then, if necessary, shaping the product intothe desired formulation.

Dry powder compositions for topical delivery to the lung by inhalationmay, for example, be presented in capsules and cartridges of for examplegelatine or blisters of for example laminated aluminium foil, for use inan inhaler or insufflator. Formulations generally comprise a powder mixfor inhalation of the salt of the invention and a suitable powder base(carrier substance) such as lactose or starch. Use of lactose ispreferred. The powder base may include additional components such aspreservatives, stabilizing agents, absorption enhancers or aerodynamicmodifier.

Each capsule or cartridge may generally contain between 0.1 μg and 9000μg of each therapeutically active ingredient. Alternatively, the activeingredient (s) may be presented without excipients.

Packaging of the formulation may be suitable for unit dose or multi-dosedelivery. In the case of multi-dose delivery, the formulation can bepre-metered or metered in use. Dry powder inhalers are thus classifiedinto three groups: (a) single dose, (b) multiple unit dose and (c) multidose devices.

For inhalers of the first type, single doses have been weighed by themanufacturer into small containers, which are mostly cartridges or hardgelatine capsules. In the case of a cartridge, the single-dose inhaleris thus composed of a cartridge containing the inhalation powder andmetering the individual dosages. The powder for inhalation ispermanently situated in the bottom of cartridge, in a reservoir with ametering slide at the base and a lid at the top. When a capsule is usedas a container, the capsule has to be taken from a separate box orcontainer and inserted into a receptacle area of the inhaler. Next, thecapsule has to be opened or perforated with pins or cutting blades inorder to allow part of the inspiratory air stream to pass through thecapsule for powder entrainment or to discharge the powder from thecapsule through these perforations by means of centrifugal force duringinhalation. After inhalation, the emptied capsule has to be removed fromthe inhaler again. Mostly, disassembling of the inhaler is necessary forinserting and removing the capsule, which is an operation that can bedifficult and burdensome for some patients.

Other drawbacks related to the use of hard gelatine capsules forinhalation powders are (a) poor protection against moisture uptake fromthe ambient air, (b) problems with opening or perforation after thecapsules have been exposed previously to extreme relative humidity,which causes fragmentation or indenture, and (c) possible inhalation ofcapsule fragments. Moreover, for a number of capsule inhalers,incomplete expulsion has been reported (e. g. Nielsen et al, 1997).

Some capsule inhalers have a magazine from which individual capsules canbe transferred to a receiving chamber, in which perforation and emptyingtakes place, as described in WO 92/03175. Other capsule inhalers haverevolving magazines with capsule chambers that can be brought in linewith the air conduit for dose discharge (e. g. WO91/02558 and GB2242134). They comprise the type of multiple unit dose inhalers togetherwith blister inhalers, which have a limited number of unit doses insupply on a disk or on a strip.

Blister inhalers provide better moisture protection of the medicamentthan capsule inhalers. Access to the powder is obtained by perforatingthe cover as well as the blister foil, or by peeling off the cover foil.When a blister strip is used instead of a disk, the number of doses canbe increased, but it is inconvenient for the patient to replace an emptystrip. Therefore, such devices are often disposable with theincorporated dose system, including the technique used to transport thestrip and open the blister pockets.

Multi-dose inhalers do not contain pre-measured quantities of the powderformulation. They consist of a relatively large container and a dosemeasuring principle that has to be operated by the patient. Thecontainer bears multiple doses that are isolated individually from thebulk of powder by volumetric displacement. Various dose measuringprinciples exist, including rotatable membranes (e. g. EP0069715) ordisks (e. g. GB 2041763; EP 0424790; DE 4239402 and EP 0674533),rotatable cylinders (e. g. EP 0166294; GB 2165159 and WO 92/09322) androtatable frustums (e. g. WO 92/00771), all having cavities which haveto be filled with powder from the container. Other multi dose deviceshave measuring plungers with a local or circumferential recess todisplace a certain volume of powder from the container to a deliverychamber or an air conduit (e. g. EP 0505321, WO 92/04068 and WO92/04928), or measuring slides such as the Genuair® devise (formerlyknown as Novolizer SD2FL) which is described in the following patentapplications: WO 97/000703, WO 03/000325 and WO2006/008027

Additional Therapeutic Agents

The salts of the present invention can also be used in combination withother drugs known to be effective in the treatment of the diseases orthe disorders indicated above. For example the salts of the presentinvention can be combined with (a) corticosteroids, or gluococorticoids(b) antihistamines (c) chemokine receptor antagonists, such as maravirocor enfuvirtide, (e) CRTH₂ antagonists, (f) leukotriene receptorantagonists, (g) JAK inhibitors such as tofacitinib or INCB018424, (h)Syk inhibitors (i) phosdiesterase IV inhibitors (j) p38 Inhibitors suchas ARRY-797, (k) PKC inhibitors such as NVP-AEB071, (l) 5-lipoxygenaseactivating protein inhibitors, such as veliflapon, (m) 5-lipoxygenaseinhibitors, (n) CYSLTR1 antagonists (o) CYSLTR2 antagonists (p) BLT1antagonists, (q) BLT2 antagonists, (r) thromboxane A2 antagonists suchas ramatroban, (s) DP1 receptor antagonists, such as laropiprant, (t)DP1 receptor agonists, such as BW-245C, (u) IP receptor agonists, suchas RO-1138452, (v) Anti-IgE, such as omalizumab, (w) IL5 antibody, suchas mepolizumab, (x) leukotriene formation inhibitors, (y) decongestants,such as ephedrine, levo-methamphetamine, naphazoline, oxymetazoline,phenylephrine, phenylpropanolamine, propylhexedrine, pseudoephedrine,synephrine or tetrahydrozoline; (z) mucolytics such as acetylcysteine,ambroxol, bromhexine, carbocisteine, domiodol, eprazinone, erdosteine,letosteine, neltenexine, sobrerol, stepronin or tiopronin; (aa)antitussives, such as dextromethorphan, (bb) analgesics such as aspirin,paracetamol, rofecoxid, celecoxib, morphine, codeine, oxycodone,hydrocodone, dihydromorphine or flupirtine; and (cc) expectorants suchantimony pentasulfide, guaiacolsulfonate, guaifenesin, potassium iodideor tyloxapol.

Accordingly, another embodiment of the invention is a combinationproduct comprising (i) at least a salt compound as defined previously,and (ii) one or more active ingredients as described above, forsimultaneous, separate or sequential use in the treatment of the humanor animal body.

A preferred embodiment of the invention is a combination product asdefined above for the treatment or prevention of pathologicalconditions, diseases and disorders associated with both β2 adrenergicreceptor and muscarinic antagonist activity, in particular wherein thepathological condition or disease is selected from asthma, acute orchronic bronchitis, emphysema, or Chronic Obstructive Pulmonary Disease(COPD), preferably asthma and COPD, as well as a method for treating asubject afflicted with a pathological condition or disease associatedwith both β2 adrenergic receptor and muscarinic antagonist activity, inparticular wherein the pathological condition or disease is as describedabove; which comprises administering to said subject an effective amountof a combination product as defined above.

As indicated above, the salts according to the invention may also beused in combination with another therapeutically active agent as definedabove.

The amount of each active which is required to achieve a therapeuticeffect will, of course, vary with the particular active, the route ofadministration, the subject under treatment, and the particular disorderor disease being treated.

The active ingredients may be administered from 1 to 6 times a day,sufficient to exhibit the desired activity. Preferably, the activeingredients are administered once or twice a day.

Examples of suitable PDE4 inhibitors that can be combined with saltcompounds of the present invention are benafentrine dimaleate,etazolate, denbufylline, rolipram, cipamfylline, zardaverine,arofylline, filaminast, tipelukast, tofimilast, piclamilast,tolafentrine, mesopram, drotaverine hydrochloride, lirimilast,roflumilast, cilomilast, oglemilast, apremilast, tetomilast, filaminast,(R)-(+)-4-[2-(3-Cyclopentyloxy-4-methoxyphenyl)-2-phenylethyl]pyridine(CDP-840),N-(3,5-Dichloro-4-pyridinyl)-2-[1-(4-fluorobenzyl)-5-hydroxy-1H-indol-3-yl]-2-oxoacetamide(GSK-842470), 9-(2-Fluorobenzyl)-N6-methyl-2-(trifluoromethyl)adenine(NCS-613), N-(3,5-Dichloro-4-pyridinyl)-8-methoxyquinoline-5-carboxamide(D-4418),3-[3-(Cyclopentyloxy)-4-methoxybenzyl]-6-(ethylamino)-8-isopropyl-3H-purinehydrochloride (V-11294A),6-[3-(N,N-Dimethylcarbamoyl)phenylsulfonyl]-4-(3-methoxyphenylamino)-8-methylquinoline-3-carboxamidehydrochloride (GSK-256066),4-[6,7-Diethoxy-2,3-bis(hydroxymethyl)naphthalen-1-yl]-1-(2-methoxyethyl)pyridin-2(1H)-one(T-440),(−)-trans-2-[3′-[3-(N-Cyclopropylcarbamoyl)-4-oxo-1,4-dihydro-1,8-naphthyridin-1-yl]-3-fluorobiphenyl-4-yl]cyclopropanecarboxylicacid (MK-0873), CDC-801, UK-500001, BLX-914,2-carbomethoxy-4-cyano-4-(3-cyclopropylmethoxy-4-difluroromethoxyphenyl)cyclohexan1-one,cis[4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-ol,CDC-801 and5(S)-[3-(Cyclopentyloxy)-4-methoxyphenyl]-3(S)-(3-methylbenzyl)piperidin-2-one(IPL-455903).

Examples of suitable corticosteroids and gluococorticoids that can becombined with salt compound of the present invention are prednisolone,methylprednisolone, dexamethasone, dexamethasone cipecilate, naflocort,deflazacort, halopredone acetate, budesonide, beclomethasonedipropionate, hydrocortisone, triamcinolone acetonide, fluocinoloneacetonide, fluocinonide, clocortolone pivalate, methylprednisoloneaceponate, dexamethasone palmitoate, tipredane, hydrocortisoneaceponate, prednicarbate, alclometasone dipropionate, halometasone,methylprednisolone suleptanate, mometasone, mometasone furoate,rimexolone, prednisolone farnesylate, ciclesonide, butixocortpropionate, RPR-106541, deprodone propionate, fluticasone, fluticasonepropionate, fluticasone furoate, halobetasol propionate, loteprednoletabonate, betamethasone butyrate propionate, flunisolide, prednisone,dexamethasone sodium phosphate, triamcinolone, betamethasone17-valerate, betamethasone, betamethasone dipropionate,21-Chloro-11beta-hydroxy-17alpha-[2-(methylsulfanyl)acetoxy]-4-pregnene-3,20-dione,Desisobutyrylciclesonide, hydrocortisone acetate, hydrocortisone sodiumsuccinate, NS-126, prednisolone sodium phosphate and hydrocortisoneprobutate, Prednisolone sodium metasulfobenzoate and clobetasolpropionate.

Examples of suitable anti-histamines that can be combined with the saltsof the invention are methapyrilene, mequitazine, azelastinehydrochloride, acrivastine, emedastine difumarate, emedastine fumarate,loratadine, cyproheptadine hydrochloride, diphenhydramine hydrochloride,doxepin hydrochloride, promethazine hydrochloride, levocabastinehydrochloride, desloratadine, cinnarizine, setastine hydrochloride,mizolastine, ebastine, cetirizine hydrochloride, epinastinehydrochloride, olopatadine hydrochloride, bepotastine besilate,triprolidine hydrochloride, rupatadine fumarate, fexofenadinehydrochloride, levocetirizine dihydrochloride, ketotifen, azatadinemaleate, dimethindene maleate, clemastine fumarate, alcaftadine,bilastine, vapitadine hydrochloride, AZD-1744, GSK-1004723D, GSK-835726or SUN-1334H.

Examples of suitable leukotriene antagonist that can be combined withthe salts of the present invention are CYSLTR1 antagonists, such asmontelukast, pranlukast or zafirlukast; or CYSLTR2 antagonists, such aspranlukast, zafirlukast or tipilukast.

Examples of suitable CRTH₂ antagonist that can be combined with thesalts of the present invention are ramatroban, AMG-009, OC-000459).

Examples of suitable Syk kinase inhibitors that can be combined with thesalts of the present invention are fosfamatinib (from Rigel), R-348(from Rigel), R-343 (from Rigel), R-112 (from Rigel), piceatannol,2-(2-Aminoethylamino)-4-[3-(trifluoromethyl)phenylamino]pyrimidine-5-carboxamide,R-091 (from Rigel),6-[5-Fluoro-2-(3,4,5-trimethoxyphenylamino)pyrimidin-4-ylamino]-2,2-dimethyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazin-3-onebenzenesulfonate (R-406 from Rigel),1-(2,4,6-Trihydroxyphenyl)-2-(4-methoxyphenyl)ethan-1-one,N-[4-[6-(Cyclobutylamino)-9H-purin-2-ylamino]phenyl]-N-methylacetamide(QAB-205 from Novartis), CI-1002 (from Pfizer), VRT-750018 (fromVertex), PRT-062607,2-[7-(3,4-Dimethoxyphenyl)imidazo[1,2-c]pyrimidin-5-ylamino]pyridine-3-carboxamidedihydrochloride (BAY-61-3606 from Bayer) and AVE-0950 (fromSanofi-Aventis).

Treatment of a Pathological Conditions or Diseases Associated with Bothβ2 Adrenergic Receptor and Muscarinic Antagonist Activity

The salts of the invention, pharmaceutical compositions and thecombinations of the invention may be used in the treatment ofpathological conditions or diseases associated with both β2 adrenergicreceptor and muscarinic antagonist activity, typically respiratorydiseases. The respiratory disease is preferably one in which the use ofbronchodilating agents is expected to have a beneficial effect, forexample asthma, acute or chronic bronchitis, emphysema, or ChronicObstructive Pulmonary Disease (COPD). Asthma or chronic obstructivepulmonary disease are more preferred.

The active compounds in the combination and the second therapeutic agentas defined above, may be administered together in the samepharmaceutical composition or in different compositions intended forseparate, simultaneous, concomitant or sequential administration by thesame or a different route.

It is contemplated that all active agents would be administered at thesame time, or very close in time. Alternatively, one or two activescould be taken in the morning and the other(s) later in the day. Or inanother scenario, one or two actives could be taken twice daily and theother(s) once daily, either at the same time as one of the twice-a-daydosing occurred, or separately. Preferably at least two, and morepreferably all, of the actives would be taken together at the same time.Preferably, at least two, and more preferably all actives would beadministered as an admixture.

The active substance compositions according to the invention arepreferably administered in the form of compositions for inhalationdelivered with the help of inhalers, especially dry powder inhalers,however, any other form or parenteral or oral application is possible.Here, the application of inhaled compositions embodies the preferredapplication form, especially in the therapy of obstructive lung diseasesor for the treatment of asthma.

The active compound(s) formulations generally contain a suitable carrierwhich may be either a propellant for MDI administration or water foradministration through a nebuliser. The formulation may compriseadditional components such as preservatives (for example, benzalkoniumchloride, potassium sorbate, benzyl alcohol); pH stabilizers (forexample, acidic agents, alkaline agents, buffer systems); isotonicstabilizers (for example, sodium chloride); surfactant and wettingagents (for example, polysorbates, sorbitan esters); and/or absorptionenhancers (for example, chitosan, hyaluronic acid, surfactants). Theformulation may also contain additives to improve the solubility ofother active compounds when mixed with the salt of the invention. Thesolubility enhancers may comprise components such as cyclodextrins,liposomes or co-solvents such as ethanol, glycerol and propylene glycol.

Additional suitable carriers for formulations of the active salts of thepresent invention can be found in Remington: The Science and Practice ofPharmacy, 20th Edition, Lippincott Williams & Wilkins, Philadelphia,Pa., 2000.

The carrier for a pharmaceutical composition in the form of a dry powderis typically chosen from starch or a pharmaceutically acceptable sugar,such as lactose or glucose. The amount of the active ingredient to thecarrier will generally vary from 0.001% to 99%.

The invention further encompasses a method of treating diseases orconditions associated with both β2 adrenergic receptor and muscarinicantagonist activity, typically respiratory diseases, such as asthma orchronic obstructive pulmonary disease, in a mammal, the methodcomprising administering to the mammal, a therapeutically effectiveamount of a salt, pharmaceutical composition or combination of theinvention. The mammal is preferably a human being.

EXAMPLES

Reagents, starting materials, and solvents were purchased fromcommercial suppliers and used as received.

Crystallizations test of salts of compounds of formula (I) with a broadrange of pharmaceutically acceptable acids (comprising among othersfumaric, succinic, sulphuric, 1-hydroxy-2-naphthoic, L-tartaric,hydrobromic, hydrochloric, oxalic, triphenylacetic, methanesulfonic,p-toluensulfonic, naphthalene-2-sulfonic, saccharin, mandelic,L-mandelic, maleic, 1S-camphor-10-sulfonic, L-malic, L-pyroglutamic andnaphthalene-1,5-disulfonic acids) in a range of differentpharmaceutically acceptable solvents (including among others acetone,ethyl acetate, isopropanol, 2-butanol, ethanol, chloroform, methanol andtetrahydrofurane or mixtures thereof) have been undertaken.

The salts from methanesulfonic, p-toluensulfonic andnaphthalene-2-sulfonic acids were obtained as solid but with a very lowcrystallinity. Salts obtained from mandelic acid and L-mandelic acid areeither oils or amorphous solids. On the other hand, the salts fromhydrochloric acid and hydrobromic acid were unstable.

Only the salts of the invention were crystalline. In addition thiscrystalline properties, salts were neither hygroscopic nor deliquescentand had a relatively high melting point allowing them to be micronizedand to have long term stability.

Particularly good methods to prepare the addition salts of the inventionare illustrated in the following examples.

The FTIR spectra were recorded using either a Bruker Alpha spectrometer,equipped with a Bruker Diamond single reflection ATR system, amid-infrared source as the excitation source and a DTGS detector, orusing a Perking Elmer, Spectrum one spectrometer, equipped with aDiamond single reflection ATR system, a mid-infrared source as theexcitation source and a DTGS detector. The spectra were acquired in 32scans at a resolution of 4 cm-1 in the range of 4000-400 cm⁻¹.

DSC analyses were recorded either in a Mettler Toledo DSC822e or using aDSC-821 Mettler-Toledo, serial number 5117423874. In the case of aMettler Toledo DSC822e equipment, samples of 1-3 mg were weighted (usinga microscale MX5, Mettler) into 40 μL aluminium crucibles with a pinholelid, and were heated, under nitrogen flow (50 mL/min), from 30 to 300°C. at a heating rate of 10° C./min. Data collection and evaluation wasdone with software STARe. In case of a DSC-821 Mettler-Toledo, serialnumber 5117423874 equipment, samples were weighed into an aluminium pan,an aluminium pinhole lid placed on top of the sample and compressed witha brass rod. Samples were equilibrated at 25° C. and heated at 10°C./min to 300° C. The instrument was calibrated using indium and zincstandards.

Thermogravimetric analyses were recorded in a Mettler Toledo SDTA851 e.Samples of 1-3 mg were weighted (using a microscale MX5, Mettler) 40 μLaluminium crucibles with a pinhole lid, and were heated at 10° C./minbetween 30 and 300° C., under nitrogen flow (50 mL/min). Data collectionand evaluation was done with software STARe.

Proton nuclear magnetic resonance analyses were recorded in a BrukerAvance 500 Ultrashield NMR spectrometer, a Varian VNMRS 600 MHz withcoldprobe and a Varian Mercury-plus 400 MHz Spectra were acquiredsolving 8-10 mg of sample in 0.5 mL of deuterated solvent.

In order to acquire a powder diffraction pattern of the obtained solid,approximately 20 mg of the non-manipulated samples were prepared instandard sample holders using foils of polyacetate.

Powder diffraction patterns were acquired on a Bruker D8 Advance Series2Theta/Theta powder diffraction system using CuKα1-radiation (1.54060 A)in transmission geometry. The system is equipped with a V{hacek over(A)}NTEC-1 single photon counting PSD, a Germanium monochromator, aninety positions auto changer sample stage, fixed divergence slits andradial soller. Programs used: Data collection with DIFFRAC plus XRDCommander V.2.4.1 and evaluation with EVA V.12.0.

Powder diffraction patterns were also performed on a Brucker X-raypowder diffractometer, model D2 Phaser with a Cu X-ray source. Themethod runs from 5 to 40 degrees 2-Theta with a 0.01 degree 2-Theta stepsize and a 0.4 second collection time at each step using a Lynxeyedetector.

Example 1 Preparation oftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate saccharinate 1.1 Preparation of free baseoftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate from hydrofluoride salt thereof

0.92 g oftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate dihydrofluoride are suspended in 10 ml ofsaturated sodium hydrogen carbonate solution. 40 ml of THF are added andthe mixture is stirred at room temperature for 10 minutes. The solventsare eliminated under vacuum (bath temperature: 40° C.) and the residueis treated with 50 ml of THF. After adding a teaspoon of Celite® anddrying with sodium sulphate, the solids are filtered. The filtrate isconcentrated to give 0.9 g of the free base.

1.2 Preparation of amorphous form oftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate saccharinate

trans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate (100 mg) was dissolved in isopropanol (6mL) at 60° C. Saccharin (50 mg) was added directly over the solution.The obtained suspension was stirred for 1 hour, cooled to roomtemperature and filtered off under vacuum. A yellowish amorphous solidwas obtained. Yield 86 mg (70%).

2.3 Preparation of Crystalline salt oftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate saccharinate from the amorphous form

The non-crystalline saccharinate salttrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate (60 mg, 0.081 mmol) was suspended inethanol (1 mL) and stirred at 70° C. for 2 hours. The suspension wasallowed to cool to a room temperature, and the obtained off-white powderwas filtered off and dried overnight under vacuum at 60° C. Yield: 50 mg(85%).

2.4 Direct Preparation of crystalline salt oftrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate saccharinate 2.4.1 Using Absolute Ethanolas a Solvent

A solution of 15.64 mg of saccharine solved in 0.544 mL of absoluteethanol is added to a hot mixture of 60.42 mg oftrans-4-((3-(5-((2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino)-methyl)-1H-benzo[d][1,2,3]triazol-1-yl)propyl)(methyl)-amino)cyclohexyl2-hydroxy-2,2-di(thiophen-2-yl)acetate in 0.302 mL of absolute ethanol.The mixture is heated at 65° C. for 4 h. After formation of the salt,the mixture is then let to cool down, filtered, washed and dried undervacuum. 56.4 mg (74.8% yield) of the title product is obtained. Ethanol96% may also be used as a solvent.

2.4.2 Using EtOH/Ethyl acetate as a Solvent

A solution of 110 mg of saccharine solved in 2 mL of a mixture EtOH96%-ethyl acetate (3:1) is added to a hot mixture of 410 mg oftrans-4-((3-(5-((2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino)methyl)-1H-benzo[d][1,2,3]triazol-1-yl)propyl)(methyl)-amino)cyclohexyl2-hydroxy-2,2-di(thiophen-2-yl)acetate in 6.5 mL of ethanol 96%. Themixture is heated at 65° C. for 3.5 h. After formation of the salt, themixture is then let to cool down, filtered, washed and dried undervacuum. 367 mg (71.9% yield).

FIG. 1 shows the powder X-ray diffraction (PXRD) pattern for thesaccharinate salt. A large number of peaks were observed thus confirmingthe crystallinity of the salt. The summary of the XRPD angles andrelative intensities are given in Table 1.

TABLE 1 Diffraction Angle (°2θ) d value (Å) Relative Intensity (%) 8.7710.08 40.3 12.35 7.16 49.9 12.92 6.85 37.8 13.35 6.63 33.8 13.85 3.3930.5 15.43 5.74 33.2 17.68 5.01 52.4 18.17 4.88 74.3 18.86 4.70 35 20.364.36 42.8 21.12 4.20 56.7 21.79 4.07 52.6 21.97 4.04 47.6 22.23 4.0054.7 22.63 3.93 34.3 24.04 3.70 30.5 24.79 3.59 59.2

FIG. 2 corresponds to the ¹H-NMR spectrum of the saccharinate salt. Itclearly shows a stoichiometry ratio of 1:1 free base/saccharin, asinferred from the comparison between the integral values of the protonscorresponding to the aromatic ring of the saccharin molecule and that ofa single proton of the hydroxyl radical of the parent structure.

¹H NMR (500 MHz, MeOD-d₄) (δ ppm): 1.3-1.6 (m), 1.95 (m, 2H), 2.13 (m,2H), 2.35-2.45 (m, 3H), 2.53 (s, 3H), 2.78-2.93 (m, 3H), 3.17 (m, 2H),4.36 (s, 2H), 4.81 (m, 1H), 4.87 (t, 2H), 5.42 (dd, 1H), 6.62 (d, 1H),7.04 (dd, 2H), 7.07 (d, 1H), 7.11 (d, 1H), 7.20 (dd, 2H), 7.32 (d, 1H),7.44 (dd, 2H), 7.71 (s, 1H), 7.75 (m, 2H), 7.84 (m, 2H), 7.91 (d, 1H),8.16 (d, 1H), 8.33 (d, 1H).

FIG. 3 shows the Thermogravimetric (TG) analysis with no significantchanges until at about 130-140° C. (significant slope and loss of mass).Prior to 140° C., and in particular at the temperature range of 80-100°C., the sample does not exhibit any change. This indicates that there isno solvent/water release, confirming thus the lack of hygroscopicity ofthe sample.

On the other hand, Differential Scanning calorimetry (DSC) only shows anintense endothermic curve with an onset at about 170° C., indicating apossible decomposition of the salt. This indicates that below 170° C.,the sample does not convert into any other polymorphs and does notsuffer any decomposition, confirming thus its high stability

Example 2 Preparation oftrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate 1.1 Preparation of free base oftrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate from hydrofluoride salt thereof

To a suspension of 125 mg (0.153 mmol) oftrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate dihydrofluoride in 7 ml of CHCl₃, 17 ml ofsaturated NaHCO₃ aqueous solution were added. The mixture was stirredduring five minutes at room temperature. The solid became an oil andCHCl₃/MeOH (10:1) solution was slowly added until dissolution wasobserved (a total volume of 28 ml was added). The phases were separatedand the aqueous phase was extracted again with CHCl₃/MeOH (10:1)solution (20 ml, 10 ml). The organic phases were combined, dried underMgSO₄, filtered and solvents were concentrated under reduced pressure toobtain 121 mg of the free base as a yellow dry foam.

1.2 Direct preparation of crystallinetrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate

108 mg of thetrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate free base (0.14 mmol) were dissolved in 4.5ml of tetrahydrofuran and 2.5 ml of ethanol were added. The solution wasfiltered through a 0.45 μm syringe filter to eliminate a very slightcloudiness. To an aliquot part of 0.7 ml of the filtrate (0.014 mmol ofthe free base), a solution of 1.8 mg of fumaric acid (0.0155 mmol, 1.11eq) in 0.37 ml of EtOH was added dropwise and shaking occasionally. Theformation of a white cloudiness started after the addition of the firstdrops of the acid solution and then the precipitate increased gradually.The mixture was allowed to stand at room temperature during 96 hours.The white solid was filtered, washed with ethanol/diethyl ether (1:1)solution and then with diethyl ether to give, after drying undernitrogen pressure, 7.5 mg of the title salt. (60.1%).

FIG. 4 shows the powder X-ray diffraction (PXRD) pattern for thefumarate salt. Some very intensive peaks were observed thus confirmingthe crystallinity of the salt. The summary of the XRPD angles andrelative intensities are given in Table 2.

TABLE 2 Diffraction Angle (°2θ) d value (Å) Relative Intensity (%) 19.654.51 7.1 21.71 4.09 97.6 24.10 3.69 29.2

FIG. 5 corresponds to the ¹H-NMR spectrum of the fumarate salt. Itclearly shows a stoichiometry ratio of 1:1 free base/fumaric acid, asinferred from the comparison between the integral values of the protonscorresponding to the fumaric acid molecule and that of a single protonof the dihydroquinolinyl radical of the parent structure

¹H-NMR (400 MHz, DMSO-d₆) δ ppm 1.32-1.41 (m, 4 H), 1.67-1.78 (m, 4 H),1.88-1.94 (m, 2 H), 2.16 (s, 3 H), 2.42-2.47 (m, 2 H), 2.54-2.59 (m, 1H), 2.74-2.80 (m, 2 H), 3.91-3.94 (m, 4 H), 3.98-4.13 (m, 2 H),4.64-4.72 (m, 1 H), 5.17 (t, 1 H), 6.46 (d, J=10.2 Hz, 1 H), 6.53 (s, 2H), 6.90-6.95 (m, 2 H), 6.95-6.99 (m, 2 H), 7.03-7.09 (m, 3 H), 7.25(br. s., 1 H), 7.30-7.35 (m, 1 H), 7.36-7.41 (m, 1 H), 7.43-7.48 (m, 2H), 7.63 (s, 1 H), 8.11 (d, J=9.8 Hz, 1 H), 10.36 (br. s., 1 H).

FIG. 6 shows the DSC analysis for the fumarate salt showing only anintense endothermic curve with a maximum at 134° C., indicating apossible fusion/decomposition of the salt. This indicates that below134° C., the sample does not convert into any other polymorphs and doesnot suffer any decomposition, confirming thus its high stability

FIG. 7 shows the TG analysis for the fumarate salt. No significantchanges are observed until about 140° C., in which the salt decomposes.

The following preparations forms are cited as composition (formulation)examples:

Composition Example 1 Formulation Example 1 Formulation for Inhalationwith a DPI

Ingredient Amounttrans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-  15 mgdihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazol-1-yl]propyl}(methyl)-amino] cyclohexylhydroxy(di-2-thienyl)acetate saccharinate (micronized) Lactose 3000 mg

Formulation Example 2 Formulation for Inhalation with a DPI

Ingredient Amounttrans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-  15 mgdihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazol-3(2H)-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate (micronized) Lactose 3000 mg

Formulation Example 3 Formulation for a MDI

Ingredient Amount trans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-10 g 1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazol-1-yl]propyl}(methyl)-amino] cyclohexylhydroxy(di-2-thienyl)acetate saccharinate (micronized)1,1,1,2,3,3,3-heptafluoro-n-propane q.s. to 200 ml

Formulation Example 4 Formulation for a MDI

Ingredient Amount trans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-10 g 1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazol-3(2H)- yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2- thienyl)acetate fumarate (micronized)1,1,1,2,3,3,3-heptafluoro-n-propane q.s. to 200 ml

1. A pharmaceutically acceptable crystalline addition salt of (i) a2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1H)-one derivative and (ii) adicarboxylic acid or a sulfimide derivative, or a pharmaceuticallyacceptable solvate thereof, wherein the2-amino-1-hydroxyethyl-8-hydroxyquinolin-2(1H)-one derivative is acompound of formula (I):

wherein: R¹, R² and R³ each are independently chosen from a hydrogenatom or a C₁₋₂ alkyl group, R⁴ is chosen from a hydrogen atom, a hydroxygroup, a hydroxymethyl group or a linear or branched C₁₋₄ alkyl group,R⁵ and R⁶ each are independently chosen from a thienyl group, a phenylgroup, a benzyl group or a C₄₋₆ cycloalkyl group, V and W each areindependently chosen from a —N—, —S— or —C(O)— group, n and m each areindependently chosen from 0, 1, 2, 3, or
 4. 2. The salt according toclaim 1, wherein R¹, R² and R³ each are independently chosen from ahydrogen atom or a methyl group.
 3. The salt according to claim 1,wherein V is chosen from a —N— or —S— group and W is chosen from —N— or—C(O)— group.
 4. The salt according to claim 1, wherein n is chosen from0 or 1, and m is chosen from 2 or
 3. 5. The salt according to claim 1,wherein both R¹ and R² are a hydrogen atom, R₃ is a methyl group, V ischosen from a —N— or —S— group, W is chosen from —N— or —C(O)— group, nis 0 and m is
 3. 6. The salt according to claim 1, wherein thedicarboxylic acid is fumaric acid or the sulfimide derivative issaccharin.
 7. The salt according to claim 1, chosen from:trans-4-[{3-[5-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-1H-1,2,3-benzotriazole-1-yl]propyl}(methyl)amino]cyclohexylhydroxy(di-2-thienyl)acetate saccharinate,trans-4-[{3-[6-({[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl]amino}methyl)-2-oxo-1,3-benzothiazole-3(2H)-yl]propyl}(methyl)-amino]cyclohexylhydroxy(di-2-thienyl)acetate fumarate, or a pharmaceutically acceptablesolvate thereof.
 8. A pharmaceutical composition comprising atherapeutically effective amount of the salt according to claim 1 and apharmaceutically acceptable carrier.
 9. The pharmaceutical compositionaccording to claim 8, wherein the composition is formulated foradministration by inhalation as a dry powder.
 10. The pharmaceuticalcomposition according to claim 8, further comprising a therapeuticallyeffective amount of at least one additional therapeutic agent.
 11. Thepharmaceutical composition according to claim 10, wherein the at leastone additional therapeutic agent is chosen from: (a) corticosteroids, orgluococorticoids, (b) antihistamines, (c) chemokine receptorantagonists, (e) CRTH₂ antagonists, (f) leukotriene receptorantagonists, (g) JAK inhibitors, (h) Syk inhibitors, (i) phosdiesteraseIV inhibitors, (j) p38 Inhibitors, (k) PKC inhibitors, (l)5-lipoxygenase activating protein inhibitors, (m) 5-lipoxygenaseinhibitors, (n) CYSLTR1 antagonists, (o) CYSLTR2 antagonists, (p) BLT1antagonists, (q) BLT2 antagonists, (r) thromboxane A2 antagonists, (s)DP1 receptor antagonists, (t) DP1 receptor agonists, (u) IP receptoragonists, (v) Anti-IgE, (w) IL5 antibody, (x) leukotriene formationinhibitors, (y) decongestants, (z) mucolytics, (aa) antitussives, (bb)analgesics, and (cc) expectorants.
 12. A combination comprising a saltaccording to claim 1 and at least one additional therapeutic agentchosen from: (a) corticosteroids, or gluococorticoids, (b)antihistamines, (c) chemokine receptor antagonists, (e) CRTH₂antagonists, (f) leukotriene receptor antagonists, (g) JAK inhibitors,(h) Syk inhibitors, (i) phosdiesterase IV inhibitors, (j) p38Inhibitors, (k) PKC inhibitors, (l) 5-lipoxygenase activating proteininhibitors, (m) 5-lipoxygenase inhibitors, (n) CYSLTR1 antagonists, (o)CYSLTR2 antagonists, (p) BLT1 antagonists, (q) BLT2 antagonists, (r)thromboxane A2 antagonists, (s) DP1 receptor antagonists, (t) DP1receptor agonists, (u) IP receptor agonists, (v) Anti-IgE, IL5 antibody,(x) leukotriene formation inhibitors, (y) decongestants, (z) mucolytics,(aa) antitussives, (bb) analgesics, and (cc) expectorants. 13.(canceled)
 14. A method for treating a subject afflicted with apathological condition or disease associated with β2 adrenergic receptoragonist and M3 muscarinic receptor antagonist activity, comprisingadministering to the subject a therapeutically effective amount of thesalt according to claim
 1. 15. The method according to claim 14, whereinthe pathological condition or disease is chosen from asthma or chronicobstructive pulmonary disease.
 16. (canceled)
 17. A method for treatinga subject afflicted with a pathological condition or disease associatedwith β2 adrenergic receptor agonist and M3 muscarinic receptorantagonist activity, comprising administering to the subject thepharmaceutical composition according to claim
 8. 18. The salt accordingto claim 1, wherein R¹ and R² are a hydrogen atom and R₃ is a methylgroup.
 19. The salt according to claim 2, wherein V is chosen from a —N—or —S— group and W is chosen from —N— or —C(O)— group.
 20. The saltaccording to claim 1, wherein n is 0 and m is
 3. 21. A method fortreating a subject afflicted with a pathological condition or diseaseassociated with β2 adrenergic receptor agonist and M3 muscarinicreceptor antagonist activity, comprising administering to the subjectthe combination according to claim 12.